Effects of different KRAS mutants and Ki67 expression on diagnosis and prognosis in lung adenocarcinoma.

Lung adenocarcinoma (LUAD) is a prevalent form of non-small cell lung cancer with a rising incidence in recent years. Understanding the mutation characteristics of LUAD is crucial for effective treatment and prediction of this disease. Among the various mutations observed in LUAD, KRAS mutations are particularly common. Different subtypes of KRAS mutations can activate the Ras signaling pathway to varying degrees, potentially influencing the pathogenesis and prognosis of LUAD. This study aims to investigate the relationship between different KRAS mutation subtypes and the pathogenesis and prognosis of LUAD. A total of 63 clinical samples of LUAD were collected for this study. The samples were analyzed using targeted gene sequencing panels to obtain sequencing data. To complement the dataset, additional clinical and sequencing data were obtained from TCGA and MSK. The analysis revealed significantly higher Ki67 immunohistochemical scores in patients with missense mutations compared to controls. Moreover, the expression level of KRAS was found to be significantly correlated with Ki67 expression. Enrichment analysis indicated that KRAS missense mutations activated the SWEET_LUNG_CANCER_KRAS_DN and CREIGHTON_ENDOCRINE_THERAPY_RESISTANCE_2 pathways. Additionally, patients with KRAS missense mutations and high Ki67 IHC scores exhibited significantly higher tumor mutational burden levels compared to other groups, which suggests they are more likely to be responsive to ICIs. Based on the data from MSK and TCGA, it was observed that patients with KRAS missense mutations had shorter survival compared to controls, and Ki67 expression level could more accurately predict patient prognosis. In conclusion, when utilizing KRAS mutations as biomarkers for the treatment and prediction of LUAD, it is important to consider the specific KRAS mutant subtypes and Ki67 expression levels. These findings contribute to a better understanding of LUAD and have implications for personalized therapeutic approaches in the management of this disease.

Clinical characteristics on low-dose high-resolution computed tomography and serum tumor markers of malignant pulmonary solid small nodules and postoperative survival analysis.

PURPOSE: To observe the clinical features of low-dose, high-resolution computed tomography (LDCT) and changes in serum tumor markers in malignant pulmonary solid small nodules (MPSSN), and to analyze the difference in survival of patients with malignant and benign PSSN 3 years after surgery. METHODS: Patients were enrolled from the thoracic surgery department of three hospitals. According to pathological diagnosis, all patients were divided into the case group (MPSSN, n=157) and the control group (BPSSN, n=75). There were no significant differences in gender, smoking habit, and family disease history. All subjects were subjected to LDCT. Four serum tumor markers (CEA, SCC, NSE, ProGRP) were examined simultaneously. Two independent sample t-tests, Mann-Whitney U rank sum test and Pearson chi-square test were used for comparsions. Two-category logistic regression was performed to analyze LDCT index and serum tumor markers levels of the two groups. ROC curve was used to evaluate the diagnostic sensitivity and specificity of relevant indicators. Kaplan-Meier method, log-rank and generalized Wilcoxon test were used to analyze the survival rate of patients after surgery. RESULTS: In univariate analysis, age, nodule size, bronchial aeration sign, bronchial truncation sign, burr sign, smooth sign and lobulated sign, SCC, NSE, and ProGRP were significantly different between two groups (p<0.05 or 0.01). In the regression analysis, there was a significant correlation between MPSSN and age (X1) [95%CI (1.272, 5.257), p=0.009], nodule size (X2) [95%CI (1.066, 2.746), p=0.041], bronchial aeration sign (X3) [95%CI (1.384, 11.425), p=0.010], bronchial truncation sign (X4) [95 %CI (1.269, 13.444), p=0.018] and burr sign (X5) [95%CI (0.054, 0.661), p=0.009], ProGRP (X10) [95%CI (1.302, 2.439), p=0.040]. The stepwise regression equation is Logistic(p)=-3.014+0.950 X1+0.064 X2+1.380 X3+1.419 X4-1.666 X5+0.263 X10. Log-rank and generalized Wilcoxon test analysis showed no difference in survival rate between the two groups (log rank p=0.271, generalized Wilcoxon, p=0.139). CONCLUSIONS: The levels of CEA, SCC, NSE and ProGRP in MPSSN were increased; age, nodule size, bronchial aeration sign, bronchial truncation sign and burr sign had predictive value for MPSSN. Patients with PSSN had better survival rates at 3 years after surgery.

Analysis of risk factors for stage I lung adenocarcinoma using low-dose high-resolution computed tomography.

Risk factors for stage I lung adenocarcinoma were analyzed using low-dose high-resolution computed tomography (CT). The patients were divided into case group (stage I lung adenocarcinoma patients) and control group (benign pulmonary nodules patients). All patients were subjected to low-dose high-resolution CT. Multiple linear regression was performed to analyze the CT imaging features of the two groups. Stage I lung adenocarcinoma patients were significantly associated with nodular site (X3, upper left lobe) [95% CI (1.796, 54.695), p=0.008], nodule type (X4) (p<0.001), nodule size (X5) [95% CI (0.614, 0.803), p<0.001], spicule sign (X7) [95% CI (0.029, 0.580), p=0.008], lobulation sign (X8) [95% CI (0.048, 0.673), p=0.011]. The stepwise regression equation is: Logistic (p) =-12.009 + 2.294X3 - 0.327X4 - 0.354X5 - 2.042X7 - 1.713X8. Risk factors of low-dose and high-resolution CT imaging for patients with stage I lung adenocarcinoma are nodular site (upper left lobe), nodule type, nodule size, spicule sign, and lobulation sign.

Marinobacter shengliensis sp. nov., a moderately halophilic bacterium isolated from oil-contaminated saline soil.

Two moderately halophilic strains, designated SL013A34A2(T) and SL013A24A, were isolated from oil-contaminated saline soil from Shengli Oilfield, eastern China. Cells were found to be Gram-staining negative, aerobic, rod-shaped with a single polar flagellum. The isolates were found to grow at 10-40 °C (optimum 35 °C), pH 6.0-9.0 (optimum pH 8.0), and NaCl concentrations of 0.5-18.0 % (w/v) (optimum 3.0-6.0 NaCl). The 16S rRNA gene sequence analysis indicated that the isolates belong to the genus Marinobacter. Strain SL013A34A2(T) shares the highest 16S rRNA gene sequence similarities with strain SL013A24A (99.3 %), followed by M. hydrocarbonoclasticus CGMCC 1.7683(T) (97.8 %), M. vinifirmus CGMCC 1.7265(T) (97.8 %), and M. excellens KMM 3809(T) (97.4 %), respectively, but low similarities (93.8-96.4 %) with type strains of the other numbers of genus Marinobacter. DNA-DNA relatedness values of strain SL013A34A2(T) with strains SL013A24A, M. hydrocarbonoclasticus CGMCC 1.7683(T), M. vinifirmus CGMCC 1.7265(T) and M. excellens KMM 3809(T) were 88.7, 29.2, 33.4 and 29.4 %, respectively. The major fatty acids of strain SL013A34A2(T) were identified as C18:1 ω9c, C16:0, C12:03-OH, C12:0, C16:1 ω9c and 10-methyl C18:0. The major respiratory quinone of strain SL013A34A2(T) was found to be ubiquinone-9, and its predominant polar lipids were identified as diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol and unidentified glycolipid. The genomic DNA G + C content was found to be 56.1 mol %. Based on the phenotypic, genetic and chemotaxonomic characteristics, these two isolates are representatives of a novel species of the genus Marinobacter, for which the name Marinobacter shengliensis sp. nov. is proposed. The type strain is SL013A34A2(T)(=LMG 27740(T) = CGMCC 1.12758(T)).

Plastorhodobacter daqingensis gen. nov., sp. nov.: a non-phototrophic bacterium isolated from Daqing Oilfield.

Two aerobic Gram staining negative, non-motile, and rod-shaped strains, DQW12E81-30(T) and DQW12E6-37-1, were isolated from an oil production mixture from Daqing Oilfield, northeastern China. Phylogenetic analysis based on the nearly complete 16S rRNA gene sequences revealed that strains DQW12E81-30(T) and DQW12E6-37-1 were members of family Rhodobacteraceae, which showed 95.6-95.9 % of 16S rRNA gene sequence similarities with Pararhodobacter aggregans DSM 18938(T), Rhodobacter veldkampii CGMCC 1.5006(T), and Roseinatronobacter thiooxidans DSM 13087(T), and lower similarities (<95.1 %) with all the left type species. Growth of strains DQW12E81-30(T) and DQW12E6-37-1 occurred at pH 7-8, 15-45 °C, and 0-4 % (w/v) of NaCl. The strains could grow both in dark and in light, but neither photosynthetic pigments nor photosynthetic reaction center gene pufM were detected in the strains. These photosynthesis-related features of the two isolates were different from those of Rhodobacter and Roseinatronobacter bacteria, but similar with those of Pararhodobacter. The genomic DNA G+C contents of strains DQW12E81-30(T) and DQW12E6-37-1 were 66.9 and 63.7 mol%, respectively. The predominant ubiquinone was Q-10 for both the strains. The major polar lipids of strain DQW12E81-30(T) were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylcholine, unidentified aminolipid, unidentified glycolipid, and unidentified phospholipid. The two strains had C18:1 ω7c, C18:0, and C18:1 ω7c 11-methyl as the major fatty acids. In addition, the strains DQW12E81-30(T) and DQW12E6-37-1 had C16:1 ω7c/C16:1 ω6c, C12:0, C14:0, C14:0 3-OH/C16:1 iso I, C10:0 3-OH, which were remarkably different from those of Pararhodobacter and Roseinatronobacter. The results of phenotypic, genotypic, and chemotaxonomic characteristics analyses indicated that strains DQW12E81-30(T) and DQW12E6-37-1 were readily different from their most phylogenetically closely related genera. Plastorhodobacter daqingensis gen. nov, sp. nov. is proposed for strains DQW12E81-30(T) and DQW12E6-37-1. The type strain is DQW12E81-30(T) (=LMG 27732(T)=CGMCC 1.12750(T)).

Halodurantibacterium flavum gen. nov., sp. nov., a non-phototrophic bacterium isolated from an oil production mixture.

Three Gram-negative bacterial strains, DQW12E6-69-1(T), DQW12E61-22-1, and DQW12E6-22-1-1, were isolated from an oil production mixture from Daqing Oilfield, northeastern China. The phylogenetic analysis based on the 16S rRNA gene sequences revealed that the three strains formed a stable cluster different from the known genus in Rhodobacteraceae of Alphaproteobacteria. In addition, they were most closely related to species in genera Pararhodobacter, Rhodobacter ,and Rhodobaca with the 16S rRNA gene sequence similarities being 95.1-95.9 %. Cells of the three strains were aerobic; they do not require salt to grow but are resistant to high salinity. They could conduct chemoorganoheterotrophic growth on various carbon sources, with non-phototrophic growth observed. The genomic DNA G+C contents of the strains DQW12E6-69-1(T), DQW12E6-22-1-1, and DQW12E61-22-1 were 63.8, 63.7, and 63.6 mol%, respectively. The predominant respiratory ubiquinone of DQW12E6-69-1(T) was Q-10, and the major fatty acids were C18:1 ω7c, C(18:0), and C(10:0) 3-OH. Photosynthetic pigments and photosynthetic reaction center gene pufM were not detected. The polar lipids were phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, unidentified glycolipid, and unidentified phospholipid. On the basis of phenotypic, genotypic, and chemotaxonomic characteristics, strains DQW12E6-69-1(T), DQW12E61-22-1, and DQW12E6-22-1-1 represent a novel genus and a novel species of the family Rhodobacteraceae. The name Halodurantibacterium flavum gen. nov., sp. nov. is proposed with strain DQW12E6-69-1(T) (=LMG 27742(T) = CGMCC 1.12756(T)) as the type strain.

Seohaeicola nanhaiensis sp. nov., a moderately halophilic bacterium isolated from the benthic sediment of South China Sea.

An aerobic, Gram-staining negative, non-motile, and rod-shaped bacterial strain, SS011A0-7#2-2(T), was isolated from the sediment of South China Sea with the depth of 1,500 m. Optimum growth occurred at pH 8.0, 30 °C, and 6 % (w/v) NaCl. Strain SS011A0-7#2-2(T) did not synthesize bacteriochlorophyll a or carotenoid, neither possess photosynthesis genes. Its genome DNA G+C content was 67.9 mol%. It contained Q-10 as the predominant ubiquinone and C18:1 ω7c (52.3 %) as the major fatty acid. The major polar lipids were phosphatidylcholine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, unidentified phospholipid, and unidentified aminolipid. The 16S rRNA gene sequence analysis revealed that it was closely related to Seohaeicola saemankumensis SD-15(T), Phaeobacter gallaeciensis BS 107(T) and Roseovarius pacificus 81-2(T) in Rhodobacteraceae, with the 16S rRNA gene sequence similarities being 96.5, 95.7, and 95.6 %, respectively. However, the phylogeny of the 16S rRNA gene sequences revealed that strain SS011A0-7#2-2(T) was a member of the genus Seohaeicola. Strain SS011A0-7#2-2(T) was moderately halophilic which was different from Seohaeicola saemankumensis SD-15(T), and it showed the enzyme activities and carbon source spectrum significantly different from Seohaeicola saemankumensis SD-15(T). As its physiological and chemotaxinomic properties were different from those of Seohaeicola saemankumensis SD-15(T), strain SS011A0-7#2-2(T) represents a novel species of the genus Seohaecola. The name Seohaeicola nanhaiensis sp. nov. is proposed, with strain SS011A0-7#2-2(T) (=LMG 27733(T) = CGMCC 1.12759(T)) as the type strain.

Glycocaulis albus sp. nov., a moderately halophilic dimorphic prosthecate bacterium isolated from petroleum-contaminated saline soil.

Two novel bacterial strains, SLG210-30A1(T) and SLG210-19A2, which shared 99.9 % 16S rRNA gene sequence similarity with each other, were isolated from petroleum-contaminated saline soil in Shengli Oilfield, eastern China. Cells were Gram-stain-negative, motile, aerobic, mesophilic and moderately halophilic. They could grow chemoheterotrophically with oxygen as an electron acceptor. Morphologically, cells were typical Caulobacteria-type dimorphic prosthecate bacteria. The genomic DNA G+C contents of strains SLG210-30A1(T) and SLG210-19A2 were 61.8 mol% and 61.6 mol% respectively. Strain SLG210-30A1(T) had Q10 as the predominant respiratory ubiquinone, and C16 : 0 (28.4 %), C17 : 0 (11.6 %), C18 : 0 (22.1 %) and C18 : 1ω7c (14.0 %) as the major cellular fatty acids. The polar lipids of the two isolates were some glycolipids, a lipid, a phospholipid, an aminoglycolipid and an aminophospholipid (all unidentified). The 16S rRNA gene sequences of strains SLG210-30A1(T) and SLG210-19A2 showed the highest similarities with Glycocaulis abyssi MCS 33(T) (99.8-99.9 %), but low sequence similarities (<94.7 %) with type strains of other members of the family Hyphomonadaceae. However, the DNA-DNA relatedness of G. abyssi MCS 33(T) to strains SLG210-30A1(T) and SLG210-19A2 was 37.4±4.4 % and 36.1±1.1 %, respectively. Based on different physiological, biochemical, and phylogenetic characteristics, strains SLG210-30A1(T) and SLG210-19A2 represent a novel species of the genus Glycocaulis. The name Glycocaulis albus is therefore proposed with strain SLG210-30A1(T) ( = LMG 27741(T) = CGMCC 1.12766(T)) as the type strain. An emended description of the genus Glycocaulis is also provided.